JP6172163B2 - 検体中のヘモグロビンA1cの免疫測定方法 - Google Patents
検体中のヘモグロビンA1cの免疫測定方法 Download PDFInfo
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- JP6172163B2 JP6172163B2 JP2014557383A JP2014557383A JP6172163B2 JP 6172163 B2 JP6172163 B2 JP 6172163B2 JP 2014557383 A JP2014557383 A JP 2014557383A JP 2014557383 A JP2014557383 A JP 2014557383A JP 6172163 B2 JP6172163 B2 JP 6172163B2
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Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
- G01N33/721—Haemoglobin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
- G01N33/721—Haemoglobin
- G01N33/723—Glycosylated haemoglobin
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
BES [N,N-Bis(2-hydroxyethyl)-2-aminoethanesulfonic acid]
MOPS [3-Morpholinopropanesulfonic acid]
TES [N-Tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid]
HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid]
DIPSO [3-[N,N-Bis(2-hydroxyethyl)amino]-2-hydroxy-1-propanesulfonic acid]
TAPSO [3-(N-tris[Hydroxymethyl]methylamino)-2-hydroxypropanesulfonic acid]
POPSO [Piperazine-1,4-bis(2-hydroxypropanesulfonic acid)]
HEPPSO [N-(Hydroxyethyl)piperazine-N'-2-hydroxypropanesulfonic acid]
EPPS [3-[4-(2-Hydroxyethyl)-1-piperazinyl]propanesulfonic acid]
Tricine [N-[Tris(hydroxymethyl)methyl]glycine]
Bicine [N,N-Bis(2-hydroxyethyl)glycine]
TAPS [N-Tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid]
機器:島津HPLCシステム
移動相:10%アセトニトリル/0.1%トリフルオロ酢酸から60%アセトニトリル/0.1%トリフルオロ酢酸への直線勾配
流 速:0.8mL/min
時間:25min
モニター:吸光度280nm
移動相:10%アセトニトリル/0.1%トリフルオロ酢酸から60%アセトニトリル/0.1%トリフルオロ酢酸への直線勾配
流 速:5mL/min
モニター:吸光度280nm
前処理が施されていない全血検体の測定系では、ラテックス試薬(ラテックス粒子を含む低浸透圧の緩衝液)中に全血を添加して赤血球の溶血及びヘモグロビンのラテックス表面への吸着を完結させる必要がある。そのため、まず、ラテックスを浮遊させる緩衝液自体に赤血球を溶血させる効果があるかどうかを確認した。
下記のサンプルについて、分光光度計Ubest-V530(JASCO)で吸光度を測定した。
No.1:HbA1c標準品(凍乾品、市販)にR1緩衝液2mLを加えて復元したもの
No.2:全血10μLをR1緩衝液2mLに添加して溶血したもの
No.3:血漿10μLをR1緩衝液2mLに溶解したもの
HbA1c標準品(凍乾品、市販)を用いて以下の2通りの標準試料(HbA1c濃度は0.0%〜14.9%)を調製し、日立7170自動分析機を用いてラテックス凝集法による測定を行ない、検量線を作成した(図2)。HbA1c特異抗体として、市販のキットにも用いられている公知の抗HbA1cモノクローナル抗体(特許文献1参照)を用いた。
標準試料1:
HbA1c標準品を検体希釈液1mLで復元し、4.8μLをサンプルとした。
標準試料2:
HbA1c標準品を検体希釈液100μLで復元し、12μLをサンプルとした。
全血検体の血球層から血球を採取して溶血させた希釈サンプルと、前処理を施されていない全血サンプルをダイレクトに免疫測定した場合の反応過程の吸光度変化を図3に示した。
HbA1c標準品(凍乾品、市販)を用いて、以下の4通りの標準試料(HbA1c濃度は0.0%〜14.9%)を調製し、2μLをサンプルとして日立7170自動分析機を用いてラテックス凝集法による測定を行ない、検量線を作成した。その検量線を図4に示す。
標準試料3:
HbA1c標準品をR1緩衝液 100μLで復元したもの。
標準試料4:
HbA1c標準品を、γグロブリン1.25mg/mL 含むR1緩衝液 100μLで復元したもの。
標準試料5:
HbA1c標準品を、γグロブリン 2.5mg/mL 含むR1緩衝液 100μLで復元したもの。
標準試料6:
HbA1c標準品をγグロブリン 5mg/mL 含むR1緩衝液 100μLで復元したもの。
Claims (3)
- ラテックス粒子と前処理が施されていない全血検体を低張液中で接触させ、次いで、ラテックス粒子上に吸着したヘモグロビンA1cと抗ヘモグロビンA1c抗体を接触させることを含む、検体中のヘモグロビンA1cの免疫測定方法。
- 前記免疫測定が凝集法により行なわれる請求項1記載の方法。
- 前記低張液が、緩衝能が中性からアルカリ側で最大となるグッドバッファーを0.02〜0.2mol/Lの濃度で含む緩衝液である請求項1又は2記載の方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2013005130 | 2013-01-16 | ||
JP2013005130 | 2013-01-16 | ||
PCT/JP2013/084905 WO2014112318A1 (ja) | 2013-01-16 | 2013-12-26 | 検体中のヘモグロビンA1cの免疫測定方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2014112318A1 JPWO2014112318A1 (ja) | 2017-01-19 |
JP6172163B2 true JP6172163B2 (ja) | 2017-08-02 |
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Application Number | Title | Priority Date | Filing Date |
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JP2014557383A Expired - Fee Related JP6172163B2 (ja) | 2013-01-16 | 2013-12-26 | 検体中のヘモグロビンA1cの免疫測定方法 |
Country Status (6)
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US (1) | US20150316541A1 (ja) |
EP (1) | EP2947458A4 (ja) |
JP (1) | JP6172163B2 (ja) |
CN (1) | CN105122060A (ja) |
TW (1) | TW201441621A (ja) |
WO (1) | WO2014112318A1 (ja) |
Families Citing this family (13)
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EP3061819B1 (en) * | 2013-10-25 | 2023-05-24 | Kikkoman Corporation | HEMOGLOBIN A1c MEASUREMENT METHOD AND MEASUREMENT KIT |
CN105891519B (zh) * | 2016-07-01 | 2018-03-30 | 安邦(厦门)生物科技有限公司 | 一种用于测定糖化血红蛋白比例的试剂盒及测定方法 |
CN107490677A (zh) * | 2017-07-21 | 2017-12-19 | 王贤俊 | 一种羧基胶乳微球与糖化血红蛋白抗体的交联组合液及其交联方法 |
EP3739337A4 (en) * | 2018-01-11 | 2021-11-24 | Toyobo Co., Ltd. | DILUTION LIQUID FOR MEASUREMENT SAMPLE, KIT AND MEASUREMENT PROCEDURE |
WO2019207724A1 (ja) * | 2018-04-26 | 2019-10-31 | 株式会社ニコン | 血液成分分離デバイス、血液成分分離方法、及び血液成分分析方法 |
JP7382071B2 (ja) | 2018-09-28 | 2023-11-16 | 積水メディカル株式会社 | 糖化ヘモグロビン(%)の測定方法 |
WO2020157375A1 (en) | 2019-01-31 | 2020-08-06 | Aidian Oy | Sampling and assay kit and method for sampling a biological sample |
US20200284807A1 (en) * | 2019-03-05 | 2020-09-10 | Victor Manneh | Saturation binding ratiometric assay |
CN111057150B (zh) * | 2019-12-30 | 2021-10-29 | 深圳开立生物医疗科技股份有限公司 | 一种乳胶微球及其应用以及糖化血红蛋白检测试剂盒 |
CN114280312B (zh) * | 2020-09-27 | 2023-09-15 | 河北特温特生物科技发展有限公司 | 一种用于免疫荧光层析检测的全血分离膜及其制备方法和应用 |
WO2023147522A1 (en) * | 2022-01-28 | 2023-08-03 | GATC Health Corp | Biomarkers for early detection of diabetes |
WO2023145915A1 (ja) * | 2022-01-31 | 2023-08-03 | 積水メディカル株式会社 | 肺サーファクタントプロテインの免疫測定方法及び免疫測定試薬 |
WO2023195347A1 (ja) * | 2022-04-04 | 2023-10-12 | デンカ株式会社 | 抗ヒトヘモグロビンβ鎖モノクローナル抗体又はその抗原結合性断片、ヒトヘモグロビン及び/又は糖化ヒトヘモグロビンを検出する方法、ヒトヘモグロビン及び/又は糖化ヒトヘモグロビンの検出キット、並びに、ペプチド |
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JP2677753B2 (ja) * | 1993-07-22 | 1997-11-17 | 株式会社エスアールエル | 凝集イムノアッセイ法 |
KR100778102B1 (ko) * | 2000-08-29 | 2007-11-27 | 가부시키가이샤 티에프비 | 재현성이 양호한 응집면역측정법 및 시약 |
JP3779885B2 (ja) * | 2001-03-26 | 2006-05-31 | 株式会社堀場製作所 | 免疫測定方法 |
JP4215462B2 (ja) * | 2002-07-26 | 2009-01-28 | 藤倉化成株式会社 | 非グリコシル化ヘモグロビン、グリコシル化ヘモグロビンに特異的に結合するモノクローナル抗体の製造方法 |
CN101529230A (zh) * | 2006-10-24 | 2009-09-09 | 皇家飞利浦电子股份有限公司 | 糖化血红蛋白的定量测定 |
US8580097B2 (en) * | 2009-04-27 | 2013-11-12 | Wako Pure Chemical Industries, Ltd. | Isotachophoresis of blood-derived samples |
EP2444803A4 (en) * | 2009-06-19 | 2012-11-14 | Infopia Co Ltd | METHOD FOR MEASURING GLYCED HEMOGLOBIN |
CN101915849B (zh) * | 2010-06-30 | 2013-06-05 | 深圳市国赛生物技术有限公司 | 一种方便加样的用于测定糖化血红蛋白百分比的检测试剂 |
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- 2013-12-26 US US14/655,277 patent/US20150316541A1/en not_active Abandoned
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JPWO2014112318A1 (ja) | 2017-01-19 |
TW201441621A (zh) | 2014-11-01 |
WO2014112318A1 (ja) | 2014-07-24 |
CN105122060A (zh) | 2015-12-02 |
EP2947458A1 (en) | 2015-11-25 |
US20150316541A1 (en) | 2015-11-05 |
EP2947458A4 (en) | 2016-08-10 |
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